SARS-CoV细胞空斑试验 : = Plaque Assay in SARS-CoV Titration
目的建立适用于生物安全三级实验室操作的SARS-CoV空斑试验方法。方法SARS-CoV10倍系列稀释接种Vero-E6细胞,用0.6%琼脂糖覆盖中性红着色(琼脂糖-中性红法)或覆盖1%甲基纤维素,4%甲醛固定,结晶紫染色(甲基纤维素-结晶紫法),空斑计数。结果琼脂糖-中性红法病毒滴度为6.14 Lg pfu/mL,甲基纤维素-结晶紫法病毒滴度为6.57 Lg pfu/mL。结论二种空斑试验方法相比,病毒滴度无明显差异。甲基纤维素-结晶紫空斑试验法形成的空斑比琼脂糖-中性红法清晰、操作简单安全、结果可长期保存。.
Objective To optimize and assess the method of plaque assay of severe acute respiratory syndrome(SARS)-associated coronavirus(SARS-CoV) for pathogenesis studies and monitor the viral titers for quantifying the amounts of SARS-CoV.Method A series of 10-fold dilutions of viruses were added into Vero-E6 cells seeded plates.The plates were overlaid with DMEM containing 1% methylcellulose 4,000 CP.The Vero-E6 cells were fixed with 4% formaldehyde,and stained with 0.5% crystal violet.Plaques were counted after the plates were air dried.Result The viral titer was 6.57 Lg pfu/mL.Clear plaques were formed in wells infected with SARS-CoV.Conclusion This procedure is useful for SARS-CoV titration and has less risk of SARS-CoV handling hazards in bio-safety laboratory.The plates can be kept for a longer time than other assays.This improved plaque formation technique can also be used and applied to measure the neutralizing antibody activities against SARS-CoV infection..
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2009-08-31 2009 |
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| Erschienen: |
2009-08-31 |
| Enthalten in: |
Zur Gesamtaufnahme - year:2009 |
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| Enthalten in: |
Zhong guo bi jiao yi xue za zhi - (2009), 08 vom: 31. Aug., Seite 65-66+93 Original Letters: Enthalten in 中国比较医学杂志 (DE-600)2994198-2 (DE-600)2994198-2 北京市 |
| Reihe: |
China Academic Journals (CAJ), A, 理工A(数学物理力学天地生) = Mathematics/ Physics/ Mechanics/ Astronomy China Academic Journals (CAJ), E, 医药卫生科技 = Medicine & Public Health |
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| Sprache: |
Chinesisch |
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| Weiterer Titel: |
Plaque Assay in SARS-CoV Titration |
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| Beteiligte Personen: |
蒋虹 [VerfasserIn] |
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| Links: |
oversea.cnki.net [lizenzpflichtig] |
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| Themen: |
中国医学科学院北京协和医学院医学实验动物研究所 |
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| Anmerkungen: |
Author info:JIANG Hong,QIAO Hong-wei,CONG Zhe,TONG Wei,WEI Qiang(Institute of Laboratory Animal Science,Chinese Academy of Medical Sciences and Peking Union Medical Collage,Beijing 100021,China) |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
CAJ312991142 |
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| 520 | |a 目的建立适用于生物安全三级实验室操作的SARS-CoV空斑试验方法。方法SARS-CoV10倍系列稀释接种Vero-E6细胞,用0.6%琼脂糖覆盖中性红着色(琼脂糖-中性红法)或覆盖1%甲基纤维素,4%甲醛固定,结晶紫染色(甲基纤维素-结晶紫法),空斑计数。结果琼脂糖-中性红法病毒滴度为6.14 Lg pfu/mL,甲基纤维素-结晶紫法病毒滴度为6.57 Lg pfu/mL。结论二种空斑试验方法相比,病毒滴度无明显差异。甲基纤维素-结晶紫空斑试验法形成的空斑比琼脂糖-中性红法清晰、操作简单安全、结果可长期保存。 | ||
| 520 | |a Objective To optimize and assess the method of plaque assay of severe acute respiratory syndrome(SARS)-associated coronavirus(SARS-CoV) for pathogenesis studies and monitor the viral titers for quantifying the amounts of SARS-CoV.Method A series of 10-fold dilutions of viruses were added into Vero-E6 cells seeded plates.The plates were overlaid with DMEM containing 1% methylcellulose 4,000 CP.The Vero-E6 cells were fixed with 4% formaldehyde,and stained with 0.5% crystal violet.Plaques were counted after the plates were air dried.Result The viral titer was 6.57 Lg pfu/mL.Clear plaques were formed in wells infected with SARS-CoV.Conclusion This procedure is useful for SARS-CoV titration and has less risk of SARS-CoV handling hazards in bio-safety laboratory.The plates can be kept for a longer time than other assays.This improved plaque formation technique can also be used and applied to measure the neutralizing antibody activities against SARS-CoV infection. | ||
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